RU2500039C1 - Method for simulating primary biliary cirrhosis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: primary biliary cirrhosis is simulated by introducing 45-50% alcoholic solution of picryl sulfonic acid 0.08-0.12 ml into rat's bulbar duodenal lumen and terminal ileum every 5-10 minutes. The primary biliary cirrhosis is diagnosed 1-1.5 months later.

EFFECT: developing the hepatic morphological changes specific for the above pathology.

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Description

The invention relates to medicine and can be used as a method of modeling primary biliary cirrhosis.

A known method of modeling liver cirrhosis by introducing chemicals to a laboratory animal, according to which a 50% solution of Sovtol-1 in olive oil is administered twice a week for a month at a rate of 0.25 ml per 100 g of body weight and 10% a solution of ethanol instead of drinking water (RF Patent No. 2197018 MKI 6 G09B 23/28, 2000). This method is taken as an analog.

There is also a method for simulating cirrhosis by subcutaneous injection of a 40% solution of carbon tetrachloride in sunflower oil at a rate of 2.0 ml / kg twice a week (RF patent No. 2410762, MKI 6 G09B 23/28, 2009). This method is adopted as a prototype. However, the accuracy of modeling primary biliary cirrhosis in this way is limited.

The aim of the invention is to improve the accuracy of modeling primary biliary cirrhosis.

The technical result is achieved by that. that 0.08-0.12 ml of a 45-50% alcohol solution of picrylsulfonic acid with an interval of 5-10 minutes is introduced into the lumen of the bulbar part of the duodenum and the terminal ileum of the rat and the model of primary biliary cirrhosis is established in 1-1.5 months .

The method is implemented as follows.

To Wistar rats weighing 220-230 g under the surgical stage of nembutal anesthesia (40-60 mg / kg), 0.08-0.12 ml of 45-50 were introduced invasively through an incision or puncture into the lumen of the bulbar part of the duodenum and terminal ileum % alcoholic solution of picrylsulfonic acid. The interval between administrations was 5-10 minutes.

After 1-1.5 months, lethargy of the animal, a decrease in appetite, and itching of the skin, mainly in the limbs and trunk, are noted, which can be judged by the appearance of bloody scratches and areas of hyperpigmentation. On the legs and buttocks, xanthelasms appear.

After the formation of primary biliary cirrhosis, a liver biopsy was performed.

3 biopsies were performed in each experimental animal, followed by preparation of 3 histological sections from each biopsy. The changes characteristic of primary biliary cirrhosis were verified: the appearance of friable lymphoid-plasma cell infiltrates in the periportal region, the penetration of lymphocytes into the epithelium of the bile ducts. The bile ducts are subject to circular destruction with the formation of epithelioid cell granulomas. In the preparations, the destruction of the bile ducts with the formation of new ducts is noted. Separate groups of hepatocytes lose contact with other cells and step necrosis occurs.

Therefore, with the introduction of picrylsulfonic acid in the claimed modes, a model of primary biliary cirrhosis is obtained.

By the end of 4 weeks, stepped necrosis, cicatricial changes in the periportal fields with a decrease in the number of bile ducts were formed.

The method is confirmed by the following examples.

Example 1

A Wistar rat weighing 220 g under the conditions of the surgical stage of nembutal anesthesia (40 mg / kg) into the lumen of the bulbar part of the duodenum along the probe and the terminal ileum was injected invasively through the incision with 0.08 ml of a 45% alcohol solution of picrylsulfonic acid with an interval of 5 min

After 1 month, lethargy of the animal, decreased appetite, itching of the skin, mainly in the limbs and trunk, were noted. Bloody combs and areas of hyperpigmentation appear in the area of increased desquamation of the epithelium. On the legs and buttocks, xanthelasms appear. Clinically, this condition of the animal was regarded as primary biliary cirrhosis.

There were 3 biopsies followed by preparation of 3 histological sections from each biopsy. The changes characteristic of primary biliary cirrhosis were revealed: the appearance of loose lymphoid-plasma cell infiltrates in the periportal region, the penetration of lymphocytes into the epithelium of the bile ducts. The bile ducts are subject to circular destruction with the formation of epithelioid cell granulomas. In the preparations, the destruction of the bile ducts with the formation of new ducts is noted. Separate groups of hepatocytes lose contact with other cells and step necrosis occurs.

Therefore, with the introduction of picrylsulfonic acid in the claimed modes, a model of primary biliary cirrhosis is obtained.

By the end of 4 weeks, stepped necrosis, cicatricial changes in the periportal fields were formed, a decrease in the number of bile ducts was noted.

Example 2

A 230 g Wistar rat under the conditions of the surgical stage of non-butal anesthesia (60 mg / kg) into the lumen of the bulbar part of the duodenum through the incision and terminal ileum through the probe, 0.12 ml of 50% alcohol solution of picrilsulfonic acid was injected invasively through a wall puncture at intervals 10 min

After 1.5 months, reduced animal motor activity is noted, the coat does not shine, it is thin, there is a decrease in appetite, and itching of the skin; the animal is forced to constantly carry out scratching movements with its front and hind limbs and tongue, bloody scratches and areas of hyperpigmentation appear. On the legs and buttocks, xanthelasms appear. Clinically, this condition of the animal was regarded as primary biliary cirrhosis.

There were 3 biopsies followed by the preparation of histological sections from each biopsy. The changes characteristic of primary biliary cirrhosis were verified: the appearance of friable lymphoid-plasma cell infiltrates in the periportal region, the penetration of lymphocytes into the epithelium of the bile ducts. The bile ducts are subject to circular destruction with the formation of epithelioid cell granulomas. In the preparations, the destruction of the bile ducts with the formation of new ducts is noted. Separate groups of hepatocytes lose contact with other cells and step necrosis occurs.

Therefore, with the introduction of picrylsulfonic acid in the claimed modes, a model of primary biliary cirrhosis is obtained.

After 1-1.5 months, 32 experimental animals showed the appearance of step necrosis, cicatricial changes in the periportal zone, and a decrease in the number of bile ducts.

Information sources

1. RF patent No. 2197018, MKI 6 G09B 23/28, 2000.

2. RF patent No. 2410762, MKI 6 G09B 23/28, 2009.

Claims (1)

A method for modeling primary biliary cirrhosis by introducing a chemical substance, characterized in that 0.08-0.12 ml of a 45-50% alcohol solution of picrilsulfonic acid are injected into the lumen of the bulbar part of the duodenum and terminal ileum of the rat with an interval of 5- 10 min, and a model of primary biliary cirrhosis is established after 1-1.5 months.